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Biotechnol Lett. 2013 Feb;35(2):279-84. doi: 10.1007/s10529-012-1076-3. Epub 2012 Oct 30.

A genome walking strategy for the identification of nucleotide sequences adjacent to known regions.

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  • 1College of Life Science, Hunan Normal University; State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha, People's Republic of China. hlw1225@hotmail.com

Abstract

To identify the transposon insertion sites in a soil actinomycete, Saccharopolyspora spinosa, a genome walking approach, termed SPTA-PCR, was developed. In SPTA-PCR, a simple procedure consisting of TA cloning and a high stringency PCR, following the single primer-mediated, randomly-primed PCR, can eliminate non-target DNA fragments and obtain target fragments specifically. Using SPTA-PCR, the DNA sequence adjacent to the highly conserved region of lectin coding gene in onion plant, Allium chinense, was also cloned.

PMID:
23108875
[PubMed - indexed for MEDLINE]
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